“The composition and abundance of cyanobacteria and their toxins, microcystins (MCs), and cylindrospermopsins (CYN) were investigated using samples collected at monthly intervals from the Amudde side of Koka Reservoir from May 2013 to April 2014. Cyanobacteria were the most abundant and persistent phytoplankton taxa with Microcystis and Cylindrospermopsis species alternately dominating the phytoplankton community of the reservoir and accounting for up to 84.3 and 11.9% of total cyanobacterial abundance, respectively. Analyses of cyanotoxins in filtered samples by HPLC-DAD and LC-MS/MS identified and quantified five variants of MCs (MC-LR, MC-YR, MC-RR, MC-dmLR, and MC-LA) in all samples, with their total concentrations ranging from 1.86 to 28.3 μg L−1 and from 1.71 to 33 μg L−1, respectively. Despite the presence and occasional abundance of Cylindrospermopsis sp., cylindrospermopsin was not detected. Redundancy analysis (RDA) showed that the environmental variables explained 82.7% of the total variance in cyanobacterial abundance and microcystin concentration. The presence of considerably high levels of MCs almost throughout the year represents a serious threat to public health and life of domestic and wild animals”.

The human health risks posed by exposure to cyanobacterial toxins such as microcystin (MC) through water and fish consumption remain poorly described. During the last two decades, coastal regions of Lake Victoria such as Nyanza Gulf (Kisumu Bay) have shown severe signs of eutrophication with blooms formed by Microcystis producing MC. In this study, the spatial variability in MC concentration in Kisumu Bay was investigated which was mostly caused by Microcystis buoyancy and wind drifting. Small fish (<6 cm) mainly composed of Rastrineobola argentea were examined for MC content by means of biological methods such as ELISA and protein phosphatase inhibition assay (PPIA) and partly by chemical-analytical methods such as LC-MS/MS. Overall, the MC content in small fish was related to the MC content observed in the seston. When comparing the MC content in the seston in relation to dry weight with the MC content in small fish the latter was found three orders of magnitude decreased. On average, the ELISA-determined MC contents exceeded the PPIA-determined MC contents by a factor of 8.2 ± 0.5 (SE) while the MC contents as determined by LC-MS/MS were close to the detection limit. Using PPIA, the MC content varied from 25–109 (mean 62 ± 7) ng/g fish dry weight in Kisumu Bay vs. 14 ± 0.8 ng MC/g in the more open water of L. Victoria at Rusinga channel. Drying the fish under the sun showed little effect on MC content, although increased humidity might indirectly favor photocatalyzed MC degradation.

The City of Salem, Oregon USA, has issued a drinking water advisory on May 29, related to the presence of cylindrospermopsin and microcystin in the water supplies. The cyanotoxins originate from the Detroit Reservoir that is used as source.

The advisory concerns infants, young children and other vulnerable individuals, stating that “children under the age of six, people with compromised immune systems, people receiving dialysis treatment, people with pre-existing liver conditions, pets, pregnant women or nursing mothers, or other sensitive populations should follow this advisory. At this time, people not on this list may continue to drink the water unless additional messaging is received.”

“To understand the uptake and processing of MC-LR in humans, the pig was chosen as an animal model. This was assessed by repeated exposure for 13 weeks of eight animals dosed daily with MC-LR at 0.04 µg/kg bw, repeated with six animals over five weeks at a dose 50 times higher at 2 µg/kg bw. An analytical method was developed for MC-LR in porcine serum and also to analyse levels of free MC-LR in harvested porcine tissues, with Lemieux Oxidation employed to determine bound MC-LR in these tissues. MC-LR was not detected in the serum of treated animals from either experiment but free MC-LR was observed in the large intestine and kidney from two animals from the higher dosed group at levels of 1.4 and 1.9 µg/kg dry weight (dw) respectively. The results indicated 50% of higher dosed animals accumulated bound MC-LR in liver tissue, averaging 26.4 µg, approximately 1.1% of the dose administered. These results point to the potential uptake and accumulation of MC-LR in human liver tissue exposed chronically to sub-acute doses.”

“Increased eutrophication of water bodies promotes cyanobacterial blooming that is hazardous due to the production of various bioactive compounds. Microcystin-LR (MCLR) is among the most widespread cyanotoxins classified as possible human carcinogen, while cylindrospermopsin (CYN) has only recently been recognized as health concern. Both cyanotoxins are genotoxic; however, the mechanisms of their action differ. They are ubiquitously present in water environment and are often detected together. Therefore, we studied genotoxic potential of the binary mixture of these cyanotoxins. Human hepatoma cells (HepG2) were exposed to a single dose of MCLR (1 μg/mL), graded doses of CYN (0.01-0.5 μg/mL), and their combinations. Comet and Cytokinesis block micronucleus assays were used to detect induction of DNA strand breaks (sb) and genomic instability, respectively, along with the transcriptional analyses of the expression of selected genes involved in xenobiotic metabolism, immediate/early cell response and DNA-damage response. MCLR induced DNA sb that were only transiently present after 4 h exposure, whereas CYN, after 24 h exposure, induced DNA sb and genomic instability. The MCLR/CYN mixture induced DNA sb after 24 h exposure, but to lesser extent as CYN alone. On the other hand, induction of genomic instability by the MCLR/CYN mixture was comparable to that induced by CYN alone. In addition, patterns of changes in the expression of selected genes induced by the MCLR/CYN mixture were not significantly different from those induced by CYN alone. Our results indicate that CYN exerts higher genotoxic potential than MCLR and that genotoxic potential of the MCLR/CYN mixture is comparable to that of CYN alone.”

The Steering Group (SG) of CYANOCOST has submitted comments to the European Commission about the inclusion of MC-LR in the proposed revision of the Drinking Water Directive (DWD). Based on the facts presented in this response the SG proposes the inclusion of all microcystins in the Drinking Water Guideline in the following manner: the reference to the parameter “microcystin-LR” in the proposed Drinking Water Directive should be replaced by “sum of all detected microcystin variants”. The parametric value should stay at 1.0 µg/l.

Microcystins (MCN), β-N-methylamino-L-alanine (BMAA) and anatoxin-a were investigated in Antarctic cyanobacterial mats collected from Ross Island and the McMurdo Ice Shelf, East Antarctica during Captain Scott’s ‘Discovery’ National Antarctic Expedition (1901–1904). Ultra-performance liquid chromatography-photodiode array detection (UPLC-PDA) and tandem mass spectrometry (MS/MS) analysis were used to quantify the cyanotoxins in seven cyanobacterial mat samples. MCNs were identified in six of the mat samples at concentrations from 0.5 to 16.1 µg g–1 dry weight. BMAA was found in one sample (528 ng g–1 dry weight, total BMAA), as well as two BMAA isomers, 2,4-diaminobutyric acid (DAB) and N-(2-aminoethyl) glycine (AEG) in six samples up to 6.56 and 6.79 μg g–1 dry weight, respectively. No anatoxin-a was detected. The findings confirm that MCNs, BMAA and BMAA isomers are preserved under dry herbarium conditions. The ‘Discovery’ cyanobacterial mat samples represent the oldest polar cyanobacterial samples found to contain cyanotoxins to date and provide new baseline data for cyanotoxins in Antarctic freshwater cyanobacterial mats from prior to human activity in Antarctica, the development of the ozone hole and current levels of climatic change.

“The cyanobacterial toxins β-methylamino-L-alanine (L-BMAA) and microcystin-LR (MC-LR; a potent liver toxin) are suspected to cause neurological disorders. Adult male C57BL/6JOlaHsd mice aged approximately 11 months were subcutaneously injected for five consecutive days with L-BMAA and microcystin-LR alone, or as a mixture. A dose-range study determined a tolerable daily dose to be ~31 µg MC-LR/kg BW/day based on survival, serum liver status enzymes, and relative liver and kidney weight. Mice tolerating the first one-two doses also tolerated the subsequent three-four doses indicating adaptation. The LD50 was 43–50 μg MC-LR/kg BW. Long-term effects (up to 10 weeks) on spatial learning and memory performance was investigated using a Barnes maze, were mice were given 30 µg MC-LR/kg BW and/or 30 mg L-BMAA/kg BW either alone or in mixture for five consecutive days. Anxiety, general locomotor activity, willingness to explore, hippocampal and peri-postrhinal cortex dependent memory was investigated after eight weeks using Open field combined with Novel location/Novel object recognition tests. Toxin exposed animals did not perform worse than controls, and MC-LR exposed animals performed somewhat better during the first Barnes maze re-test session. MC-LR exposed mice rapidly lost up to ~5% body weight, but regained weight from day eight.”

The European Commission has issued a proposal for a Directive on the Quality of Water Intended for Human Consumption. The proposal is a recast of Directive 98/83/EC, which was amended in 2003, 2009 and 2015.

Microcystin-LR is included in the proposal as a “Chemical Parameter”, with a parametric value of 1μg/L, along with other organic compounds that are added in the list (Bisphenol A, Haloacetic acids, Nonylphenol, PFAS). Performance criteria for determination of MC-LR are specified as 30% uncertainty at the parametric value, while methods should comply to the requirements of ISO 17025.

Inclusion of MC-LR in the proposed Directive is a result of increased occurrence of microcystins in European waters and beyond, as well as of increased awareness of the health risks and hazards associated with toxic cyanobacteria. CYANOCOST has significant contributions in this field, including two books on chemical analysis of cyanotoxins and molecular detection of toxigenic cyanobacteria and many joint parers on cyanotoxin research. Since 2012, CYANOCOST has also contributed in raising awareness of cyanotoxins in Europe; in this sense, the proposed Directive is rewarding of the vast amount of voluntarily work done within the Network and highlights the important societal impact of COST Actions.

The proposed Directive is under public consultation till 2 April 2018. You can download the related documents and submit your feedback here.